响应面法优化末水坛紫菜蛋白酶解工艺及其酶解液抗氧化活性研究
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摘要
为实现末水坛紫菜(Porphyra haitanensis)的高值化利用,研究了末水坛紫菜的蛋白酶解工艺及其酶解液的抗氧化活性。以酶解产物水解度和还原力为指标,分别采用单因素和响应面优化实验筛选出最适蛋白酶和最佳酶解工艺参数;通过测定酶解液还原力对1,1-二苯基-三硝基苯肼(DPPH)自由基、羟自由基(·OH)和超氧阴离子自由基(O_2~-·)的清除作用,研究了最高水解度下的酶解液的抗氧化性活性。结果表明,中性蛋白酶是6种蛋白酶中的最适用酶;最佳酶解条件为:底物质量浓度35 g·L~(-1)、加酶量31 200 U·g~(-1)、温度45℃、pH 7.6、酶解时间5 h,在此条件下坛紫菜水解度达31.37%;酶解液还原力为2.2,对DPPH、·OH和O_2~-·自由基清除率分别为56.26%、85.84%和72.73%。结果表明,中性蛋白酶可以有效水解末水坛紫菜,水解后的酶解产物具有较好的抗氧化能力和应用前景。
In order to achieve high-value utilization of abandoned Porphyra haitanensis, we studied the enzymatic hydrolysis technique and antioxidant activity of P. haitanensis protein. Single factor test and response surface optimization test were used to screen the optimum protease and enzymolysis parameters based on the indices of hydrolysis degree and reducing power. The antioxidant activity of the hydrolysate was studied by determining the reducing power of hydrolysate and scavenging ability of hydrolysate on the free radicals DPPH, ·OH and O_2~-·. The results show that neutral protease was the optimal enzyme among the six enzymes. The optimal conditions for the hydrolysis of P. haitanensis proteins were as follows: substrate concentration 35 g·L~(-1), enzyme dosage31 200 U·g~(-1), hydrolysis temperature 45 ℃, pH 7.6, hydrolysis time 5 h. Under these conditions, the hydrolysis degree was 31.37%and the reductive power was 2.2. The scavenging rates for DPPH, ·OH and O_2~-· were 56.26%, 85.84% and 72.74%, respectively,which indicates that P. haitanensis can be hydrolyzed by neutral protease effectively, and its hydrolysate has good antioxidant activity and application prospects.
In order to achieve high-value utilization of abandoned Porphyra haitanensis, we studied the enzymatic hydrolysis technique and antioxidant activity of P. haitanensis protein. Single factor test and response surface optimization test were used to screen the optimum protease and enzymolysis parameters based on the indices of hydrolysis degree and reducing power. The antioxidant activity of the hydrolysate was studied by determining the reducing power of hydrolysate and scavenging ability of hydrolysate on the free radicals DPPH, ·OH and O_2~-·. The results show that neutral protease was the optimal enzyme among the six enzymes. The optimal conditions for the hydrolysis of P. haitanensis proteins were as follows: substrate concentration 35 g·L~(-1), enzyme dosage31 200 U·g~(-1), hydrolysis temperature 45 ℃, pH 7.6, hydrolysis time 5 h. Under these conditions, the hydrolysis degree was 31.37%and the reductive power was 2.2. The scavenging rates for DPPH, ·OH and O_2~-· were 56.26%, 85.84% and 72.74%, respectively,which indicates that P. haitanensis can be hydrolyzed by neutral protease effectively, and its hydrolysate has good antioxidant activity and application prospects.
引文
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[31]MACHU L,MISURCOVA L,AMBROZOVA J V,et al.Phenolic content and antioxidant capacity in algal food products[J].Molecules(Basel,Switzerland),2015,20(1):1118-1133.
[32]ZHAO Z Y,ZHANG Q,LI Y F,et al.Optimization of ultrasound extraction of Alisma orientalis polysaccharides by response surface methodology and their antioxidant activities[J].Carbohyd Polym,2015,119:101-109.
[33]陈利梅.超声波辅助双水相提取条斑紫菜黄酮类物质及其抗氧化活性研究[J].食品科学,2012,33(4):41-46.
[34]龚明,黄宽,叶丽芝,等.枳木具子解酒方护肝活性成分提取工艺优化及药效学研究[J].中国临床药理学与治疗学,2016,21(2):125-129.
[35]彭新颜,孔保华,熊幼翎.由碱性蛋白酶制备的乳清蛋白水解物抗氧化活性的研究[J].中国乳品工业,2008(4):8-12,47.
[2]王贤,张希春,陈昭华.热水法和微波法提取末水坛紫菜多糖的研究[J].集美大学学报(自然科学版),2014,19(6):409-415.
[3]陈美珍,徐景燕,潘群文,等.末水残次坛紫菜的营养成分及多糖组成分析[J].食品科学,2011,32(20):230-234.
[4]吴凤娜,贾秀春,李迎秋.紫菜蛋白的研究进展[J].山东食品发酵,2013(4):44-47.
[5]高丹丹,祁高展,郭鹏辉.响应面法优化风味蛋白酶水解棉籽蛋白工艺[J].食品工业科技,2014,35(17):222-226.
[6]李媛媛,孙方达,刘骞,等.水解度对玉米蛋白水解物抗氧化性和乳化性影响的研究[J].食品工业,2016,37(8):201-207.
[7]谢飞,曹纯洁,陈美珍,等.响应面试验优化末水坛紫菜多糖除蛋白工艺及其抗氧化活性[J].食品科学,2016,37(22):77-84.
[8]朱晓连,陈华,蔡冰娜,等.具有结合胆酸盐作用卵形鲳鲹蛋白酶解物的制备和分子量分布研究[J].南方水产科学,2017,13(2):101-108.
[9]张意静.食品分析技术[M].北京:中国轻工业出版社,2001:186-194.
[10]宁正祥.食品成分分析手册[M].北京:中国轻工业出版社,1998:120-121.
[11]WU P P,MA G Z,LI N,et al.Investigation of in vitro and in vivo antioxidant activities of flavonoids rich extract from the berries of Rhodomyrtus tomentosa(Ait.)Hassk[J].Food Chem,2015,173:194-202.
[12]NAJAFIAN L,BABJI A S.Production of bioactive peptides using enzymatic hydrolysis and identification antioxidative peptides from patin(Pangasius sutchi)sarcoplasmic protein hydolysate[J].J Funct Foods,2014,9:280-289.
[13]ZHANG Y F,DUAN X,ZHUANG Y L.Purification and characterization of novel antioxidant peptides from enzymatic hydrolysates of tilapia(Oreochromis niloticus)skin gelatin[J].Peptides,2012,38(1):13-21.
[14]XIE Z J,HUANG J R,XU X M,et al.Antioxidant activity of peptides isolated from alfalfa leaf protein hydrolysate[J].Food Chem,2008,111(2):370-376.
[15]梁晓芳,牟建楼,严超,等.扇贝酶解条件的优化及其抗氧化性的研究[J].中国调味品,2018,43(1):61-66.
[16]KLOMPONG V,BENJAKUL S,KANTACHOTE D A,et al.Comparative study on antioxidative activity of yellow stripe trevally protein hydrolysate produced from Alcalase and Flavourzyme[J].Int J Food Sci Technol,2008,43(6):1019-1026.
[17]李银娟.低苦味大豆抗氧化肽的制备及应用[D].广州:华南理工大学,2013:1-65.
[18]马涛,张海欣,马永庆,等.三文鱼小清蛋白定向酶解模型及致敏活性分析[J].食品工业,2017,38(2):179-183.
[19]WANG B,LI L,CHI C F,et al.Purification and characterisation of a novel antioxidant peptide derived from blue mussel(Mytilus edulis)protein hydrolysate[J].Food Chem,2013,138(2/3):1713-1719.
[20]张风,夏旭,周爱梅,等.虾头虾壳蛋白质酶解制备抗氧化肽的研究[J].南方水产科学,2015,11(6):79-87.
[21]ROSSINI K,NORENA C P,CLADERA-OLIVERA F,et al.Casein peptides with inhibitory activity on lipid oxidation in beef homogenates and mechanically deboned poultry meat[J].LWT-Food Sci Technol,2009,42(4):862-867.
[22]周丽丽,吴跃梅,王素英.响应面法优化酶解藻蓝蛋白制备抗肥胖肽[J].中国食品学报,2017,17(5):99-107.
[23]林伟锋.可控酶解从海洋鱼蛋白中制备生物活性肽的研究[D].广州:华南理工大学,2003:1-134.
[24]滕超,王中月,李秀婷,等.生产高纯度低聚木糖的酶筛选及其水解条件优化[J].中国食品学报,2018,18(6):104-111.
[25]张杨,胡磊,汪少芸,等.响应面优化酶解法制备蒲公英籽蛋白抗氧化肽工艺[J].食品工业科技,2016,37(5):258-262.
[26]DONG S Y,ZENG M Y,WANG D F,et al.Antioxidant and biochemical properties of protein hydrolysates prepared from silver carp(Hypophthalmichthys molitrix)[J].Food Chem,2008,107(4):1485-1493.
[27]呂小京,操德群,徐年军.海洋微藻微拟球藻抗氧化肽酶解法制备和响应面工艺优化[J].食品科学,2018,39(6):183-188.
[28]DOBLADO-MALDONADO A F,JANSSEN F,COMAND S V,et al.A response surface analysis of the aqueous leaching of amylase from maize starch[J].Food Hydrocolloid,2017,63:265-272.
[29]LIN H M,DENG S G,HUANG S B.Antioxidant activities of ferrous-chelating peptides isolated from five types of low-value fish protein hydrolysates[J].J Food Biochem,2014,38(6):627-633.
[30]CHANG C Y,WU K C,CHIANG S H.Antioxidant properties and protein compositions of porcine haemoglobin hydrolysates[J].Food Chem,2007,100(4):1537-1543.
[31]MACHU L,MISURCOVA L,AMBROZOVA J V,et al.Phenolic content and antioxidant capacity in algal food products[J].Molecules(Basel,Switzerland),2015,20(1):1118-1133.
[32]ZHAO Z Y,ZHANG Q,LI Y F,et al.Optimization of ultrasound extraction of Alisma orientalis polysaccharides by response surface methodology and their antioxidant activities[J].Carbohyd Polym,2015,119:101-109.
[33]陈利梅.超声波辅助双水相提取条斑紫菜黄酮类物质及其抗氧化活性研究[J].食品科学,2012,33(4):41-46.
[34]龚明,黄宽,叶丽芝,等.枳木具子解酒方护肝活性成分提取工艺优化及药效学研究[J].中国临床药理学与治疗学,2016,21(2):125-129.
[35]彭新颜,孔保华,熊幼翎.由碱性蛋白酶制备的乳清蛋白水解物抗氧化活性的研究[J].中国乳品工业,2008(4):8-12,47.